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Biological control of field infections of nematode parasites of young sheep with Duddingtonia flagrans and effects of spore intake on efficacy.

A field study was undertaken to determine the effects of feeding Duddingtonia flagrans to young Merino sheep on pasture. A total of 60 mixed sex lambs 4-5 months old were divided into six even groups on the basis of liveweight. On Monday to Friday, each week for 6 months, three groups were offered barley grains on which D. flagrans had been cultured while the other three groups remained untreated. Every 4 weeks liveweights were recorded and faecal samples collected for nematode egg count estimation. Feeding D. flagrans reduced faecal egg counts and tended to improve liveweight gains, but considerable differences were observed between groups within treatment. These differences are thought to result from variations between the groups in consumption of the treated barley with the "best" consumers showing the greater effects of treatment.

Animals↗

Lung granulomatous response induced by infection with the intestinal nematode Nippostrongylus brasiliensis is suppressed in mast cell-deficient Ws/Ws rats.

Certain nematode infections induce eosinophil infiltration and granulomatous responses in the lungs. To examine the role of mast cells in the development of lung lesions, normal +/+ and genetically mast cell-deficient Ws/Ws rats were infected with the nematode Nippostrongylus brasiliensis. In +/+ rats, numbers of eosinophils in bronchoalveolar lavage fluid (BALF) increased significantly 3-7 days after infection, and granulomatous responses composed of histiocytes/ macrophages and multinucleate giant cells were triggered in the lungs 3-14 days after infection. Challenge infection, which was carried out on day 28 after primary infection, induced much higher levels of granulomatous response than after primary infection, suggesting that the response is mediated at least in part by an immunological mechanism. In Ws/Ws rats, both the eosinophil percentage in BALF and the size of the granulomas in the lungs were significantly smaller than in +/+ rats after primary as well as after challenge infection. The amount of rat mast cell protease (RMCP) II in +/+ rat BALF was increased 1 day after primary infection and more significantly after challenge infection, suggesting that lung mucosal mast cells were activated more markedly after the challenge infection. In Ws/Ws rats, RMCP II was undetectable throughout the observation period. The time course of nematode migration in the lungs did not differ in +/+ and Ws/Ws rats. These results suggest that mast cell activation might be relevant to eosinophil infiltration and granulomatous response in the lungs, although the responses do not affect lung migration of the nematode.

Animals↗

Interleukin-5 deficient mice exhibit impaired host defence against challenge Trichinella spiralis infections.

Enteric nematode infections are characterized by both peripheral and tissue eosinophilia. The cytokine interleukin (IL)-5 is considered a critical factor in the proliferation and recruitment of eosinophils, however, studies suggest it plays little role in host defence, at least during primary Trichinella spiralis infections. Less is known concerning its role in host defence or in the inflammatory response that develops against challenge infections with the same parasite. We examined these questions by infecting IL-5 deficient and wild-type mice, with T. spiralis parasites. Both strains expelled the primary infection by day 21. Forty days after the primary infection, we challenged the mice with a second T. spiralis infection and counted tissue eosinophils and worms in the intestine. While wild-type mice developed a large tissue eosinophilia, IL-5 deficient mice showed little increase in eosinophil numbers within the intestine. Throughout the challenge infection, significantly larger worm burdens were recovered from IL-5 deficient mice, and worm expulsion was also significantly slower (day 21) compared to wild-type mice (day 14). Thus, unlike in a primary infection, IL-5 is not only essential for the onset of intestinal eosinophilia, but also makes a significant contribution to enteric host defence during challenge T. spiralis infections.

Animals↗

Contribution of 5-HT2A receptor in nematode infection-induced murine intestinal smooth muscle hypercontractility.

BACKGROUND & AIMS: Enteric nematode infection induces a smooth muscle hypercontractility that depends on interleukin (IL)-4 and IL-13 activation of the signal transducer and activator of transcription (STAT) 6. Serotonin (5-HT) is involved in the physiologic regulation of gut function. The present study investigated the contribution of 5-HT and its receptors in nematode-induced intestinal smooth muscle hypercontractility. METHODS: Mice were infected with Nippostrongylus brasiliensis (N brasiliensis) or Heligmosomoides polygyrus (H polygyrus) or injected intravenously with IL-13. Segments of jejunum were suspended in organ baths, and smooth muscle responses to 5-HT were determined in the presence or absence of specific 5-HT antagonists. IL-4, IL-13, and 5-HT receptor messenger RNA expressions were determined by real-time quantitative polymerase chain reaction. RESULTS: 5-HT evoked a modest contraction of smooth muscle in wild-type (WT) mice that was unaltered by the 5-HT2A antagonist ketanserin. N brasiliensis infection induced a smooth muscle hypercontractility to 5-HT that was abolished by 5-HT(2A) antagonists but not by other 5-HT antagonists. Infection-induced up-regulation of 5-HT2A expression was correlated with the smooth muscle hypercontractility to 5-HT. The infection-induced up-regulation of 5-HT2A in WT mice was observed also in IL-4(-/-) mice but was not seen in IL-13(-/-) or STAT6(-/-) mice. In addition, smooth muscle responses to 5-HT and 5-HT2A expression in WT mice were also enhanced by IL-13 or H polygyrus infection. CONCLUSIONS: These data show that 5-HT2A is one of the molecules downstream from STAT6 activation that mediates changes in smooth muscle function. 5-HT2A represents a novel therapeutic target for modulating immune-mediated effects on intestinal motility.

Animals↗

Internal parasite levels and response to anthelmintic treatment by beef cows and calves.

Albendazole (methyl 5-propylthio-1 H-benzimidazol-2-yl carbamate) was used as an anthelmintic in a 3-yr study involving 578 beef cows and 438 nursing calves. Infection levels for nematodes, coccidia, and tapeworm were relatively low throughout the 3-yr period. Eggs per gram of feces in cows and calves were lower (P less than .01) 2 wk posttreatment but were not different 5 mo later, when calves were weaned. Cow weight gain, rate and time of conception, and adjusted calf weaning weights were not affected significantly by deworming of either cows or calves. Level of nematode infection measured as eggs per gram of feces was higher (P less than .01) in younger cows than in mature cows. Although deworming with Albendazole lowered (P less than .01) nematode infection levels, no responses were observed in cow or calf performance.

Age Factors↗

Effects of the nematode Gyrinicola batrachiensis on development, gut morphology, and fermentation in bullfrog tadpoles (Rana catesbeiana): a novel mutualism.

We describe a novel mutualism between bullfrog tadpoles (Rana catesbeiana) and a tadpole-specific gastrointestinal nematode (Gyrinicola batrachiensis). Groups of tadpoles were inoculated with viable or nonviable nematode eggs, and development, morphology, and gut fermentation activity were compared between nematode-infected and uninfected tadpoles. Nematode infection accelerated tadpole development; the mean time to metamorphosis was 16 d shorter and the range of times to metamorphosis was narrower in nematode-infected tadpoles than in uninfected tadpoles. At metamorphosis, infected and uninfected bullfrogs did not differ in body size or condition. Colon width, wet mass of colon contents, and concentrations of most fermentation byproducts (short-chain fatty acids: SCFAs) in the hindgut were greater in infected tadpoles. Furthermore, in vitro fermentation yields for all SCFAs combined were over twice as high in infected tadpoles than in uninfected tadpoles. One explanation for accelerated development in infected tadpoles is the altered hindgut fermentation associated with the nematodes. Energetic contributions of fermentation were estimated to be 20% and 9% of the total daily energy requirement for infected and uninfected tadpoles, respectively. Infection by G. batrachiensis nematodes potentially confers major ecological and evolutionary advantages to R. catesbeiana tadpoles. The mutualism between these species broadens our understanding of the taxonomic diversity and physiological contributions of fermentative gut symbionts and suggests that nematodes inhabiting the gut regions of other ectothermic herbivores might have beneficial effects in those hosts.

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Efficacy of a topical formulation of eprinomectin against endoparasites of cattle in New Zealand.

Two controlled studies involving 24 cattle were conducted in New Zealand to determine the efficacy of a topical, non-flammable formulation of eprinomectin against induced and naturally acquired nematode infections. In Trial 1, nematode infections were induced on Day -5 with third-stage larvae of Cooperia spp., Haemonchus contortus, Ostertagia ostertagi and Trichostrongvlus colubriformis so that the nematodes would be at the fourth larval stage when the cattle were treated. In Trial 2, cattle had naturally acquired nematode infections as determined by faecal nematode egg counts and larval cultures. The cattle were allocated on Day 0 (Trial 1) or Day 6 (Trial 2) on a stratified random basis according to bodyweight to one of two treatments: untreated control or eprinomectin (0.5% w/v) applied topically at 1 ml/10 kg bodyweight. Necropsies were undertaken on Days 14 and 15 and total nematode counts were done. In Trial 1, cattle treated with eprinomectin had significantly (p < 0.05) fewer Cooperia spp. and O. ostertagi than the controls. Larvae of H. contortus and T. colubriformis did not establish. In Trial 2, cattle treated with eprinomectin had significantly (p < 0.05) fewer of the following parasites than the controls: Haemonchus spp. (adult), Cooperia surnabada (adult), C. oncophora (adult), Cooperia spp. (L,), Ostertagia lyrata (adult), O. ostertagi (adult), Oesophagostomum spp. (adult), T. avei (adult and L1) and Trichuris spp. (adult). Reductions of 100% were observed for Capilfaria spp. (adult), D. viviparus (adult and L,), and Nematodirus helvetianus (adult), but these were not statistically significant (p > 0.05) because four or fewer control animals were infected with these parasites. In Trial 2, efficacies of greater than 99% were observed against all species for which moderate to high burdens occurred in the untreated controls. These findings indicate that eprinomectin in a topical formulation is a highly effective nematocide in cattle.

Journal Article↗

Natural infection by gastrointestinal and bronchopulmonary nematodes in mouflons (Ovis musimon) and their response to netobimin treatment.

Gastrointestinal and bronchopulmonary nematode infections and the efficacy of netobimin (Hapasil) were analyzed by way of fecal examination in 10 female mouflons (Ovis musimon), in central Spain, February 1993. Before treatment all 10 mouflons had Trichostrongylus axei, Teladorsagia circumcincta and Marshallagia spp.; sic had Nematodirus spp., two had Trichuris sp., one had Capillaria sp., seven had bronchopulmonary Dictyocaulus filaria and 10 mouflons had protostrongylid lungworms (Muellerius capillaris, Protostrongylus rufescens, Cystocaulus ocreatus or Neostrongylus linearis). Netobimin (7.5 mg/kg) was 100% effective against T. axei, T. circumcincta, Marshallagia spp., and D. filaria infections whereas one animal continued eliminating Nematodirus spp. eggs. The drug also was effective against Capillaria spp. but not against Trichuris spp. or protostrongylid infections.

Animals↗

Bromodeoxyuridine labeling studies on the proliferation of intestinal mucosal mast cells in normal and athymic rats.

The proliferation of mucosal-type mast cells (MMC) in rat small intestine was studied using a bromodeoxyuridine (BrdU)-labeling method. After 24-h cumulative injections of BrdU into adult SD rats, 3.5% of MMC were labeled, while only 0.3% and 0.1% of mast cells were labeled in back skin and ear respectively. From the results, it was concluded that MMC division occurred more than 10 times as frequently as the division of skin mast cells. Similar results were obtained in athymic adult rats (F344/N Jcl-rnu) in which the number of MMC was similar to that in heterologous animals. Thus, thymic factor(s) or T cells may not have an important role in MMC division in normal states. When SD rats were infected with Nippostrongylus brasiliensis, vigorous proliferation of MMC was brought about 13 to 15 days after infection. At that period, 40% of MMC were labeled by a single injection of BrdU, and 85% of MMC were labeled by 9-h cumulative injections of BrdU, with the result that most MMC rapidly proliferated in the intestinal mucosa during this period. Mitotic figures of MMC were sometimes observed. On the contrary, hyperplasia of MMC was not observed in athymic rats infected with nematodes. Therefore, MMC hyperplasia after nematode infection is dependent on thymic factor or T cells, and its mechanism is different from that of MMC division in normal states, in which thymic factor(s) or T cells are not essential.

Animals↗

Intestinal nematode parasites, cytokines and effector mechanisms.

Laboratory models of intestinal nematode infection have played an important role in developing our understanding of the immune mechanisms that operate against infectious agents. The type of helper T cell response that develops following infection with intestinal nematode parasites is critical to the outcome of infection. The early events that mediate polarisation of the helper T cell subsets towards either Th1 or Th2 during intestinal nematode infection are not well characterised, but it is likely that multiple factors influence the induction of a Th1 or Th2 type response, just as multiple effector mechanisms are involved in worm expulsion. Costimulatory molecules have been shown to be important in driving T helper cell development down a specific pathway as has the immediate cytokine environment during T cell activation. If helper T cells of the Th2 type gain ascendancy then a protective immune response ensues, mediated by Th2 type cytokines and the effector mechanisms they control. In contrast, if an inappropriate Th1 type response predominates the ability to expel infection is compromised. Equally important is the observation that multiple potential effector mechanisms are stimulated by nematode infection, with a unique combination operating against the parasite depending on nematode species and its life cycle stage. Despite the close association between intestinal nematode infection and the generation of eosinophilia, mastocytosis and IgE it has been difficult to consistently demonstrate a role for these effector cells/molecules in resistance to nematode parasites, although mast cells are clearly important in some cases. It therefore seems that, in general, less classical Th2 controlled effector mechanisms, which remain poorly defined, are probably important in resistance to nematode parasites. Thus, our understanding of both the induction and effector phases remains incomplete and will remain an intense area of interest in the coming years.

Animals↗

Effect of herd management on the contamination of night holding areas (correos) and infections with gastrointestinal nematodes of N'Dama cattle in The Gambia.

The densities of infective nematode larvae (L3/m2) in the night holding places (locally called "correos") of 2 traditionally kept N'Dama herds were estimated at weekly intervals throughout an entire rainy season. Herd 1 moved correos every 3 weeks whereas herd 2 remained in the same area for most of the rainy season. Removal to a new correo was invariably accompanied by a drastic drop of L3/m2. Conversely, L3/m2 increased rapidly up to values of more than 1,000 when the herds used the same night holding place for more than 3 weeks. Calves kept in herds with frequent changes of the correo showed significantly lower nematode egg counts and higher growth rates during the rainy season, combined with a reduced weight loss during the following dry season. The results of this study indicate that a regular frequent change of the correo is an effective method of reducing nematode infection risk and increasing calf growth and that this might be a sustainable part of an integrated strategic programme to control gastrointestinal nematode infections wherever correos are used.

Animal Husbandry↗

Critical role for signal transducer and activator of transcription factor 6 in mediating intestinal muscle hypercontractility and worm expulsion in Trichinella spiralis-infected mice.

Intestinal nematode infections in rats or mice are accompanied by intestinal muscle hyper contractility that may contribute to parasite expulsion from the gut. Previous studies demonstrated that both the expulsion of nematode parasites and the associated muscle hyper contractility are dependent on CD4(+) T helper cells. Nevertheless, the precise immunological mechanism underlying changes in intestinal muscle function remains to be determined. In this study, we investigated the role of interleukin 4 (IL-4) and signal transducer and activator of transcription factor 6 (STAT6) in the development of intestinal muscle hypercontractility and worm expulsion by infecting IL-4 and STAT6-deficient mice with Trichinella spiralis. Worm expulsion was almost normal in IL-4-deficient mice but substantially delayed in STAT6-deficient mice. Consistent with delayed worm expulsion, we also observed a marked attenuation of carbachol-induced muscle contraction in STAT6-deficient mice but only a moderate decrease in muscle hypercontractility in IL-4-deficient mice. In addition, we also observed severe impairment of T helper type 2 cytokine responses and intestinal mucosal mastocytosis in STAT6-deficient mice, although some degree of intestinal tissue eosinophilia was evident in these animals. These results are consistent with the hypothesis that STAT6-dependent changes in intestinal muscle function contribute to host protection in nematode infection.

Animals↗

Comparison of naturally acquired parasite burdens among Florida native, Rambouillet and crossbreed ewes.

The response to natural infection by gastrointestinal nematodes was evaluated in three groups of lambing mature ewes: 16 Florida Native; 21 Rambouillet; and 18 1/2 Florida Native x1/2 Rambouillet (F1 generation). Fecal egg counts (FEC), packed cell volume (PCV) and plasma protein levels were determined periodically through a one-year period. The highest FEC was recorded in Rambouillet ewes, with most of the means higher than 1000 eggs per gram (EPG). Florida Native and F1 ewes showed similar trends in FEC, with most of the means below 500 EPG. The highest mean PCV values were recorded in Florida Native ewes, followed in decreasing order by F1 and Rambouillet ewes. F1 ewes showed the lowest plasma protein levels and Florida Natives the highest. Fecal cultures revealed the highest percentages of Haemonchus larvae in Rambouillet ewes (>64%) and the lowest percentages in Florida Native ewes. Florida Native and F1 ewes were more resistant to gastrointestinal nematode infections than were Rambouillet ewes. High negative correlation coefficients between FEC x PCV and FEC x plasma protein levels coincided with peaks in mean FEC of Rambouillet (May and July), Florida Native and F1 ewes (October). This study showed that crossbreeding Rambouillet and Florida Native sheep resulted in the production of F1 ewes resistant against gastrointestinal nematode infections.

Animals↗

Functional relationships between sensory nerve fibers and mast cells of dura mater in normal and inflammatory conditions.

In this study, we have characterized the phenotype of mast cells in rat dura mater and their topological and functional relationships with C-fibers in normal and inflammatory conditions. Three mast cell populations with different size, morphology and localization were characterized by their content of specific neutral serine proteases. They showed immunoreactivity corresponding to rat mast cell protease I, rat mast cell protease II, or both proteases. Using confocal microscopy, all three mast cell types were observed in close apposition (distance less than 100 nm) to calcitonin gene-related peptide- and substance P-immunoreactive nerve fibers in both controls and rats infected with the nematode Nippostrongylus brasiliensis. After nematode infection or neonatal treatment with capsaicin, a large increase in the number of rat mast cell protease II-immunoreactive mast cells was found within dura mater segments (+1478% and +596%, respectively), without concomitant changes of rat mast cell protease I- or rat mast cell protease I/II-immunoreactive mast cells. Under both these conditions, the increase in mast cell number was accompanied by a significant increase in rat mast cell protease II level within tissue extracts (+281% after nematode infection and +36% after capsaicin treatment). The functional interaction of mast cells with sensory nerve fibers in the dura mater was assessed by evaluating [3H]histamine synthesis after administration of L-[3H]histidine, an index of mast cell activity. The H3 receptor agonist (R)-alpha-methylhistamine (15 mg/kg, i.p.) had no effect, but administration of the H3 receptor antagonist, thioperamide (10 mg/kg, i.p.), resulted in a significant increase of [3H]histamine synthesis (+62%). This effect was reduced in neonatal capsaicin-treated rats, but not completely suppressed (+35%), very likely because of partial denervation, as assessed by monitoring calcitonin gene-related peptide immunoreactivity. It is concluded that, in the dura mater, as in peripheral tissues, sensory nerve fibers and mast cells actively synthesizing and releasing histamine form a short inhibitory feedback loop involving prejunctional H3 receptors that could regulate the release of pro-inflammatory mediators, thus limiting the extent of inflammatory reactions.

Animals↗